Effects of a Novel Pathogenesis-Related Class 10 (PR-10) Protein from Crotalaria pallida Roots with Papain Inhibitory Activity against Root-Knot Nematode Meloidogyne incognita

A novel pathogenesis-related class 10 (PR-10) protein with papain inhibitory activity, named CpPRI, was purified from Crotalaria pallida roots by ammonium sulfate precipitation followed by three reverse-phase high-performance liquid chromatographies (HPLCs). CpPRI is made up of a single polypeptide...

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Veröffentlicht in:Journal of agricultural and food chemistry 2010-04, Vol.58 (7), p.4145-4152
Hauptverfasser: Andrade, Lúcia Betânia da S, Oliveira, Adeliana S, Ribeiro, Jannison K. C, Kiyota, Sumika, Vasconcelos, Ilka Maria, de Oliveira, José Tadeu Abreu, de Sales, Maurício P
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Sprache:eng
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Zusammenfassung:A novel pathogenesis-related class 10 (PR-10) protein with papain inhibitory activity, named CpPRI, was purified from Crotalaria pallida roots by ammonium sulfate precipitation followed by three reverse-phase high-performance liquid chromatographies (HPLCs). CpPRI is made up of a single polypeptide chain with a M r of 15 kDa, as determined by sodium dodecyl sulfate−polyacrylamide gel electrophoresis (SDS−PAGE). This protein exhibited a K i value of 1.8 × 10−9 M and operates via a noncompetitive inhibition mechanism. The alignment of the N-terminal amino acid sequence of CpPRI with other proteins revealed its identity with PR-10 proteins. CpPRI acts against digestive proteinase from root-knot nematode Meloidogyne incognita and demonstrated nematostatic and nematicide effects on this parasite in bioassays. In a localization study, fluorescein-5-isothiocyanate (FITC)−CpPRI was observed to internalize and diffuse over the entire J2 body after 6 h of incubation. This fact could explain the natural tolerance of this plant species to nematodes.
ISSN:0021-8561
1520-5118
DOI:10.1021/jf9044556